FI112211B - Process for the preparation of therapeutically useful 15-deoxy pergualin analogs and intermediates used in the process - Google Patents

Abstract

The present invention relates to compounds of structure related to that of 15-deoxyspergualine, as new industrial products. These new compounds correspond to the formula: <IMAGE> in which: - n is equal to 6 or 8, - A represents a single bond, CH2, CHF, CH(OH), CH(OCH3), CH2NH or CH2O, and their addition salts. These new compounds are useful as immunosuppressants.

Description

112211

The present invention relates to novel compounds structurally related to 15-deoxyspergualin and relates to a process for their preparation. The compounds may be used in therapy as immunosuppressive agents.

It is known that 15-deoxyspergualin is a derivative of spergualin, spergualin being an antibiotic isolated from a culture of Bacillus laterosporus. Initial studies of 15-deoxy-spergualin revealed its antitumor activity, later its activity of 15 has been investigated primarily in the immunosuppression region.

Reference may be made in particular to G. Dickneite, "15-Deoxyspergualin: From Cy-20 Tostasis to Immunosuppression", Behring Inst.Mitt., No.

82: 231-239 (1988); G.Dickneite, "The Influence of (±) - 15-Deoxyspergualin on Experimental Transplantation and Its Immunopharmacological Mode of Action," Behring Inst.

Mitt, no. 80, 93-102 (1986) and K. Nemoto, "Deoxyspergu-25 alin in Lethal murine graft-versus-host disease", Trans-Plantation vol. 51, 712-715, no. 3, March 1991.

In spite of its actual activity in the immunosuppression region, 15-deoxyspergualin does not have satisfactory chemical stability. As a result, * * 'attempts have been made to obtain more stable derivatives, in particular by stopping the off-hydroxyglycine residue of deoxyspergualin with various ex- or ω-amino acids.

* _ * 35 In this context, reference may be made in particular to »» *: R. Nishizawa, "Synthesis and Biological Activity of Sper *": Analogues of Gualin, "J. Antibiotics 1988, 42 (11), 1629-1643 and EP-A -0 105 193.

2 112211

New 15-deoxyspergualin analogs have now been obtained which are structurally different from the products disclosed in EP-A-0 105 193 and which are chemically stable and exhibit greater activity in immunosuppression than the known products.

A notable difference between the products made and known according to the invention in terms of chemical structure is the inversion of a CO-NH bond linking a guanidine hexyl or 10 guanidinooctyl residue to a central amino acid.

The 15-deoxyspergualiline analogue compounds prepared according to the invention include: 15 (i) Compounds of formula I nh o ^ (CH 2) n il / / (CH 2> A \ / <CH 2> 3 \ I * J, 20 "

(D

wherein n is 6 or 8 and A is a single bond, CH 2, CH (OH), CHF, CH (OCH 3), CH 2 NH or CH 2 O, or ♦ t ♦ ♦, 25 (ii) the addition salts thereof.

The invention thus relates to a process for the preparation of the compounds of the formula I and: to their addition salts, wherein the compound of the formula V

R

: / 1 'NH o o p \ (CH,) U 11 ^ (CH,),. ___ (CH2) 3 ^ R1 r, (vii) 35>; · where n and A have the same meaning as above and Rx is amii- * * **. deprotection, especially I · t is used to replace R3 with H.

3, 112211

The immunosuppressive agent of the formula I or its non-toxic addition salt can be used for the preparation of a medicament for use in therapy for the suppression of immune diseases.

5

The compound of formula I, or a non-toxic addition salt thereof, may also be used in the preparation of medicaments for the treatment of malaria.

It goes without saying that in such uses, the therapeutically effective amount of the active agent is used.

Addition salts are acid addition salts obtained by reaction of a mineral acid or an organic acid with a compound of the formula I. Preferred mineral acids for salt formation are hydrochloric, hydrobromic, sulfuric and phosphoric acids. Preferred organic acids for salt formation are fumaric, maleic, methanesulfonic, oxalic, citric and trifluoroacetic acids.

20

The compounds of formula I may be prepared by methods known per se using conventional reaction mechanisms such as amide bond formation and, in particular, by application of methods known in the art of peptide chemistry.

25 ...... .........................

As stated above, the preparation process of the invention includes deprotection of the compound of formula VII.

In practice, this means that each protecting group R 1; which will be replaced by a hydrogen atom is a type of oxycarbonyl, known in the peptide syntheses, which is temporarily protected by unsaturated amino functions. The residues which are read below with their usual abbreviations may be mentioned

* I I

suitable protecting groups for this purpose: 4 112211

Adoc = adamantyloxycarbonyl

Aoc = tert.-amyloxycarbonyl

Boc = tert-butoxycarbonyl (alternative name: (1,1-dimethylethoxy) carbonyl) 5 Fmoc = 9-fluorenylmethoxycarbonyl

Foe = furfuryloxycarbonyl

Iboc = isobornyloxycarbonyl Z = benzyloxycarbonyl Z (p-Cl) = p-chlorobenzyloxycarbonyl 10 X (p-OMe) = p-methoxybenzyloxycarbonyl Of these groups, hereinafter referred to as amino protecting groups, the preferred R 1 group according to the invention is the Boc group.

15

Processes for the preparation of a compound of formula I or an addition salt thereof include: Process variant A, which comprises: (i) Compound of formula II

1 'NH

R ^ (CH) ^

:! n ^ NH

Wherein n is 6 or 8 and R 2 is an amino protecting group, in particular a / (1,1-dimethylethoxy) carbonyl group, is reacted; with an acid or an acid chloride of formula III oo 7 {III) wherein X is a chlorine atom or a group OH, A is a single 1 'bond, CH 2, CHF, CH (OCH 2 C 6 H 5) or CH (OCH 3) and R 2 is 7 35 linear or branched C 1 -C 4 alkyl group or phenyl> -methyl group, in an organic solvent (especially a chlorinated solvent such as dichloromethane or chloroform) a carboxy group activator (especially a carbodiimide such as 1,3-disyclide). -hexylcarbodiimide), and in the presence of a nucleophilic agent (especially 1-hydroxybenzotriazole) at a temperature of 0 ° C to about 40 ° C, with a molar ratio of the compounds of formula II to about 1 mol of compound of formula III , to give a compound of formula IV, 10 R 11 'is R 1 / <CH 2> ^ 11 R where R 1f R 2 and n are as defined above and A is a single bond, CH 2, CHF, CH (OCH 2 C 6 H 5) or CH (OCH 3). ,

(ii) saponifying the resulting compound of formula IV in an organic solvent in the presence of a strong base to give a compound of formula V

1 'NH O O

(v)> wherein R2 and n have the same meaning as above and A is a single bond, CH2, CHF, CH (OCH2C6H5) or CH (OCH3), »t | *> '(Iii) condensing the resulting compound of formula V *? * * with an amine of formula VI: * · *: 30 u / <ch2) 3 \ •. H2N ^ nh (vi)

• · I

· ... · R, • I »• I ♦ ii. ' wherein R 1 has the same meaning as above, under the conditions which

': * Are the same as in step (i) above, formula VII

To obtain the compound of 35 · tl »6 112211

Rl 'NH O 0 R1.N ^' s ^ NH ^ '(CH2) n \ ^ (CH2> 4 \ .- ^ <CH2> 3 \ / R1

• N NH- ^ n ^ NH A ^ NH * ^ N · "^ NH

R, (VII) 5 wherein R 1 and n are as defined above and A is a single bond, CH 2 / CHF, CH (OCH 2 C 5 H 5) or CH (OCH 3), (iv) a compound of formula VII wherein A is CH-10 (OCH 2 ), optionally deprotecting by catalytic hydrogenation to provide a compound of formula VII wherein A is a CH (OH) group, (v) deprotecting a compound of formula VII obtained in step (iii) or (iv) wherein A is a single bond , CH2, CHF, CH (OH) or CH (OCH3), in particular by reaction with a strong acid, such as trifluoroacetic acid, to remove the protecting group R1f with a compound of formula I wherein A is a single bond, CH2, CHF, CH (OH) or CH (OCH 3) to provide the addition salt, and, where appropriate, the compound of formula I is obtained in the form of the free base by reaction with a strong base; with and after said free base gives *. 25 daan other addition salts, * '·

• * 1 · i * I

In the step: (i) a compound of formula VI

• · I

· · ·: '* 30 ^ (CH2) 4 ^ (CH,),. ^ r, h2n z 3 ^ -nh (VI)

· ... * I

R1 • tl I I | wherein R 3 is a protecting group as set forth above [especially lit ·, (1,1-dimethylethoxy) carbonyl] is reacted * <· · ;;; 35 with an acid or acid chloride of formula III 7112211

° O

.A. (111)

5 X A

wherein X is a chlorine atom or an OH group, A is a single bond, CH2, CH (OCH2C6H5), CH (OCH3) or CHF and R2 is a linear or branched C1-C3 alkyl group or a phenylmethyl group, in an organic solvent ( in particular in a chlorinated solvent such as dichloromethane or chloroform) in the presence of a carboxy group activator (especially carbodiimide such as 1,3-dicyclohexylcarbodiimide) and in the presence of a nucleophilic substance (especially 1-hydroxybenzotriazole) 0 ° C to about 40 ° C, with a molar ratio of compounds of about 1 mole of the compound of Formula VI / about 1 mole of the compound of Formula III to give a compound of Formula VIII at 20 ° C? X ^ <CH2> 4 \ ^ <CH2> 3 \

^ Ο- ^ ΝΑ ^ ΝϊΤ 2 AlH

:: i R! (VIII) • wherein R 3 and R 2 are as defined above and A is one; 25 single bond, CH2, CH (OCH2CsH5), CH (OCH3) or CHF group, | (Ii) the resulting compound of formula VIII is saponified in an organic solvent in the presence of a strong base to give a compound of formula IX.

HO ^ NH

r, ™ · ;;; Wherein Rx has the same meaning as above and A is a single bond, CH2, CH (OCH2C6H5), CH (OCH3), or CHF, the compound of formula IX formed is condensed according to formula II with an amine

'NH

5 V ^ KH ^ Vn ^ (II where n is 6 or 8 and R x is as defined above under conditions similar to Step-10a (i) above) to give a compound of formula VII (Vii) wherein R 1 and n are as defined above and A is a single bond, CH 2 -, CH (OCH 2 C 6 H 5) -. or a CH (OCH 3) or CHF group, 20 (iv) of a compound of formula VII wherein A is CH- (OCH 2 C 6 H 5), if necessary deprotected to provide a compound of formula VII wherein A is CH ( OH) group, * · *: 25 (v) of the compound of formula VII formed i *: deprotected by reaction with a strong acid (especially trifluoroacetic acid), a compound of formula I wherein A is a single bond, CH 2 -, CH (OH) , CH (OCH 3) or CHF group to give the addition salt, and 30 * (vi) said compound of formula I is obtained, if necessary, in the form of the free base by reaction with a strong base: and thereafter, from said free base, other addition salts are obtained, 35 "* Process variant C, which comprises: (i) reacting a base of formula X with a base of formula X; Where Rx is a protecting group as described above [especially (1,1-dimethylethoxy) carbonyl] and n is 6 or 8, the NH2-terminal end is acylated with chloroformate or symmetric carbonate [especially bis (4-nitro) -phenyl) carbonate] in an inert solvent at room temperature (15-25 ° C), (ii) the compound formed is aminolysed with the amine of formula VI, ie - (CH2 → 4 → (CH2 → 3 → - 1) H2N NH (VI) R1 wherein Rx is as defined above to give 20r of a compound of formula VII,

'NH O O

'· R1 A CH2> A. J-L (CH2 ___ (CH-,) ·, - R.

R, (VII) 25 wherein R j and n are as defined above and A is *. The CH2NH group, (iii) the resulting compound of formula VII is deprotected by reaction with a strong acid (especially trifluoroacetic acid) to give the addition salt of a compound of formula I wherein A is CH2NH, and (vi) said the compound of the invention is obtained, if necessary, in the form of the free base by reaction with a strong base · 35 followed by the addition of said free base with other addition salts, and 112211 it | Process variant D, which process comprises: (i) Compound VI of formula VI (CH2 ^. (CH2) 3 / -R1 h2n2 ^ NH (VI) 5 i, wherein Rx is an amino protecting group as described above [especially (1, 1-dimethylethoxy) carbonyl], is reacted with a carbonate of formula III 'at 10 ° C to 11 (HI'-) C6 H5 ° wherein R2 is a linear or branched C1-C4 alkyl group or a phenylmethyl group, in an inert organic solvent (especially aromatic in a solvent such as toluene) at the reflux temperature of the reaction mixture, at a molar ratio of compounds of about 1 mole of the compound of formula VI to about 1 mole of the compound of formula III 'to afford the compound of formula VIII. <ch2) 3 \ ^

iD VV ^ NH ^ NH

I (VIII) · R1 '* * * * * * * * wherein Rx and R2 are as defined above and A is .., CH2O group, the compound of formula VIII formed soap in an organic solvent in the presence of a strong base to give the compound of formula IX. wherein Rx has the same meaning as above and A is CH2O, (iii) the compound of formula IX formed is condensed with an amine of formula II.

V

1 NH

Wherein R 1 is as defined above and n is 6 or 8, in an organic solvent (particularly a chlorinated solvent such as dichloromethane or in chloroform) in the presence of a carboxy group activator (in particular a carbodiimide such as, for example, 1,3-dicyclohexylcarbodiimide-20-din) and in the presence of a nucleophilic substance (especially 1-hydroxybenzotriazole) at a temperature from 0 ° C to about 40 ° C. being about 1 mole of a compound of Formula IX / about 1 mole of a compound of Formula II, to provide 25 of a compound of Formula VII: R NH): · NH NHO R, A · AA ^ - (CH2) i ^ -, (CH,), ^ R, V 30 ", (VII> where R x and n are as defined above and A is CH 2 O, I (iv) deprotecting the compound of formula VII by reaction with a strong acid (especially trifluoroacetic acid). »· J 35 pon), the addition salt of the compound of formula I wherein A is CH 2 O and 12,112,211 (v), if necessary, said compound of formula I is obtained in the form of the free base by reaction with a strong base followed by the addition of said free base with other addition salts.

5

The compound of formula VI wherein R 2 is a (1,1-dimethylethoxy) carbonyl group can be obtained by the method of Raymond J. Bergeron, "Total Synthesis of (+) - 15-Deoxyspergualin", J. Org.Chem. 1987, 52, 1700-1703.

10

The compounds of formula IV can also be obtained by formation and subsequent aminolysis of the mixed anhydride: the acid of formula III is reacted with chloroformate, in particular isobutyl chloroformate, in the presence of one equivalent of a tertiary base, in particular N-methylmorpholine, at about -30 ° C for about 0.5 hours and then the base of formula II is added to the reaction mixture.

20

Formation of the amide bond can also be accomplished by methods known to those skilled in the art, in particular by acylation of the appropriate amine with inorganic acid chloride. in a solvent in the presence of a strong organic base.

Intermediates of Formula VII wherein Rx is an amino protecting group, especially of the oxycarbonyl type, n is 6 or 8 and A is a single bond, CH2, CH (OH), CH (OCH3) -, CH2NH-, CH2O, CH (OCH2C5H5) or CHF are new compounds and are within the scope of the invention.

* I

2 »3»

The invention will be more readily understood from the description of the following Examples and the pharmacological results obtained by comparing 5 to 6 compounds of the invention with compounds known in the art. The nomenclature used herein is those recommended by Chemical Abstracts: According to this nomenclature, a diester of the tert-butylethylalkane-13112211 diolate type is (1,1-dimethyl-ethyl) -ethylalkanedioate.

PREPARATIVE EXAMPLE I

5 Bis (1,1-dimethylyl) - [[(6-aminohexyl) iminoylmethylene] bis-carbamate 17.23 g (0.148 mol) of hexane-1,6-diamine are added at room temperature with stirring to a solution of 10 43 g (0.148 mol) of N, N'-bis (tert-butoxycarbonyl) -5-methylisothiourea in 300 ml of tetrahydrofuran. The re-reaction mixture is stirred for 16 hours. The solvent is evaporated, and the residue obtained is chromatographed on silica, eluting with CHCl3 / ethanol (3/1, v / v) and then with a mixture of ethyl acetate, methanol and 32% aqueous ammonia (6/3 / vol). 0.1, v / v / v), 19.7 g (37% yield) of a yellow oil are obtained.

1 HNMR (CDCl 3): 1.25-1.60 (m, 28H), 2.7 (t, 2H), 3.5 (q, 2H), 8.3 (t, 1H), 11.5 ( s, 1H).

*

The following product was obtained using an analogous procedure: • Bis (1,1-dimethyl-methyl) - [[(8-amino-octyl) -imino] -methylene] bis-carbamate. 1 H NMR (CDCl 3): 1.3-1.7 (m, 32H), 2.7 (t, 2H), 3.4 (q, 2H), 8.3 (t, 1H), 11.5 ( s, lH).

»

PREPARATIVE EXAMPLE II

1- (1,1-Dimethylethyl) -14-ethyl-6 - [(1,1-dimethylethoxybenzyl) -12-oxo-2,6,11-triazatetradecanedioate 0.82 g (4 · 10'3 moles) 1,3-dicyclohexylcarbodiimide ···. is added to a solution cooled to 0 ° C and containing 0.53 g (4 x 10'3 moles) of ethyl malonate in 20 ml. * 35 anhydrous chloroform. After stirring for 0.5 h, at 0 ° C, a solution of 1.04 g (3.10-3 mol) of 1,1-dimethylethyl-10-amino-6- [ (1,1-di-14112211 methylethoxy) carbonyl] -2,6-diazadecanoate in 5 ml of anhydrous chloroform. The mixture is stirred for 5 hours at room temperature and 1.06 g (8.10-3 mol) of ethylmalonate and 1.64 g (8.10-3 mol) of 1,3-dicyclohexyl-5-lycarbodiimide are added. The mixture is stirred for 1 hour and the solvent is evaporated under reduced pressure. The resulting pasty residue is chromatographed on silica, eluting with ethyl acetate / hexane (1/1, v / v) followed by ethyl acetate, to give 0.95 g (69% 10%) of the expected product as an oil.

1 H NMR (CDCl 3): 1.25 (t, 3H), 1.40-1.70 (m, 24H), 3.10-3.35 (m, 10H), 4.3 (q, 2H).

PREPARATIVE EXAMPLE II

1- (1,1-Dimethyl-methyl) -6 - [(1,1-dimethylethoxy) carbonyl] -12-oxo-2,6,11-triazatetradecanedioate 0.95 g (2.07110 moles) ), the product obtained in Preparation Example II is dissolved in a mixture of 20 ml of 1 N sodium hydroxide and 20 ml of dimethoxyethane (1/1, v / v). The reaction mixture is stirred for 15 minutes at room temperature, reduced to one-third volume, then acidified with 1N hydrochloric acid to pH 2-3. It is then extracted twice with 50 ml of chloro. 25 Formia. The organic phases are evaporated under reduced pressure, after which the residue obtained is chromatographed! silica, eluting with ethyl acetate / methanol (3/1, v / v) followed by methanol, gives 0.75 g (84% yield) of the expected product as an oil.

1 H NMR (Dimethylsulfoxide-d 6): 1.40 (s, 18H), 1.55 (m, 6H), 1 1. 2.90 (m, 4H), 3.15 (m, 6H) ).

* · · · * · »15 112211

PREPARATIVE EXAMPLE IV

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] - 12,14-dioxo-2/4, 11,15,20,24-Hexaza-pentacos-2-ene-5 dioate 0.46 g (2.32-10-3 moles) 1,3-dicyclohexylcarbodiimide and 0.0155 g (0.1 103 mol) of 1-hydroxybenzotriazole dihydrate are added with stirring to a solution of 10 g of the product of Preparation III at 0 ° C in 30 ml of anhydrous chloroform. After stirring the reaction mixture for 0.5 h, 0.42 g (1.16-10. 3 moles) of bis- (1,1-dimethylethyl) - [[(6-aminohexyl)] are added dropwise at 0 ° C. imino] methylene-15 ni] biscarbamate (obtained by the method of Preparation Example I above). The reaction mixture is again stirred at 0 ° C for 1.5 hours, after which 0.23 g (1.16-10.3 mol) of 1,3-dicyclohexylcarbodiimide are added and stirring is continued for 24 hours at room temperature. The solvent 20 is evaporated under reduced pressure and the resulting residue is chromatographed on silica using ethyl acetate / hexane (1/1, v / v) as the eluent followed by ethyl acetate and finally ethyl acetate / methanol. (9/1, v / v) gives 0.7 g (73% yield) of the expected product as an oil.

1 H NMR (CDCl 3): 1.3-1.7 (m, 50H), 3.1-3.3 (m, 12H), 3.4 (q, V 2H), 4.8 and 5.3 (broad s, 1H), 6.8 and 7.15 (broad s, 2H), 8.3 (t, 1H), 11.5 (s, 1H).

EXAMPLE 1 N- [4 - [[3- (amino) propyl] amino] butyl] -N '- [6 - [(amino - iminomethyl) amino] hexyl] propanediamide tris (tri-

I I I

, ···. fluoroacetate) • · 0.7 g (0.9 · 10 ~ 3 moles) of the product obtained in Preparation Example IV * are dissolved in 10 ml of trifluoroacetic acid and 10 ml of anhydrous dichloromethane. The reaction mixture is stirred for 24 hours at room temperature, after which the solvent is evaporated under reduced pressure. The residue obtained is taken up in 150 ml of distilled water and then lyophilized. The residue is purified by MPLC (medium-5 pressure liquid chromatography) reverse phase (RP18 silica) eluting with water / acetonitrile / trifluoroacetic acid (7/2/1 v / v / v) to give 0.43 g ( 66% yield) of a highly hygroscopic solid.

1 HNMR (Dimethylsulfoxide-d 6): 1.2-1.6 (m, 12H), 1.9 (m, 2H), 2.9-3.1 (m, 14H), 7.2 (broad s, 4H), 7.7 (t, 1H), 8 (m, 5H), 8.7 (broad s, 2H).

13CNMR (dimethylsulfoxide-d6): 22.5; 23.4; 25.4; 25.6; 15.7; 28.0; 28.5; 35.8; 37.6; 38.2; 40.3; 43.0; 43.5; 46.1; 156.6; 166.9 (2C).

PREPARATIVE EXAMPLE V

1- (1,1-Dimethylethyl) -16-ethyl-3 - [[(1,1-dimethylethoxy) carbonyl] amino] -14-oxo-2,4,13-triazahexadec-2-enidioate »* 1,96 g (75% yield) of the expected product are obtained in the form of an oil, following the procedure of Preparation IV. 25 and starting from 1.32 g (10.10-3 mol) of ethyl malonate *, and 2 g (5.18-103 mol) of bis (1,1-dimethylethyl) -, [[(8-amino) octyl) imino] methylene] biscarbamate.

1 H NMR (CDCl 3): 1.25-1.70 (m, 33H), 3.25 (q, 2H), 3.30 (s, 2H), 3.40 (q, 2H), 4.2 (q, 2H), 7.25 (s, 1H), 8.3 (s, 1H), δ 11.5 (s, 1H).

> i i i · 1 1 · * »• · · {» »i» · »17 112211

PREPARATIVE EXAMPLE VI

1- (1,1-Dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -14-oxo-2,4,13-triazahexadec-2-enidioate 5 1.92 g (100% yield) of the expected product is obtained as an oil following the procedure of Preparation Example III starting from 1.96 g (3.92-103 mol) of the compound obtained in Preparation Example V.

1 H NMR (CDCl 3): 1.25-1.70 (m, 30H), 3.27-3.39 (m, 6H), 7.25 (s, 1H), 8.40 (s, 1H).

PREPARATIVE EXAMPLE VII

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -22 - [(1,1-dimethylethoxy) carbonyl] - 14,16-dioxo-2,4 , 13,17,22,26-Hexaza-azaheptacos-2-ene diioate 0.41 g (13% yield) of the expected product is obtained as an oil, following the procedure of Preparation IV and starting from 1.85 g (3). , 92 · 10'3 moles) of the product obtained in Preparation VI and 1.35 g (3.92 * 10'3 moles) of 1,1-dimethylethyl-10-amino-6 - [(1, 1-dimethyl-ethoxy) carbonyl] -2,6-diazadecanoate.

3 H NMR (CDCl 3): 1.30-1.70 (m, 54H), 3.1-3.5 (m, 14H), 4.8 and 5.3 (broad s, 1H), 6.7 and 7.1 (broad s, 2H), 8.3 (t, 1H), λ 11.5 (s, 1H).

•

Example 2 30 N- (4 - [[3- (Amino) propyl] amino] butyl 3-N '- [8- ((aminoiminomethyl) amino] octyl] propanediamide Tri (tri Fluoroacetate) ) 1 ·

! f I

0.24 g (65% yield) of the expected product is obtained as an oil, following the procedure of Example 1 and starting from 0.41 g (0.51-103 moles) of the product of Preparation VII.

18 112211 1HNMR (dimethylsulfoxide-d6). 1.2-1.6 (in, 16H), 1.9 (m, 2H), 2.9-3 / 1 (m, 14H), 7.2 (broad S, 4H), 7.7 (t, 1H), 8 (m, 5H), 8.7 (broad s, 2H).

13 C NMR (dimethylsulfoxide-d 6): 22.9; 23.7; 26.0; 26.1; 26.3; 28.4; 28.5; 28.6; 28.9; 36.2; 37.9; 38.6; 40.7; 43.3; 43.8; 46.4; 156.8; 166.7; 166.9.

PREPARATIVE EXAMPLE VIII

10 1- (1,1-Dimethylethyl) -14-methyl-3 - [[(1,1-dimethylethoxy) carbonyl] amino] -13-methoxy-12-oxo-2,4,11-tri-azatetradec -2-enidioate 4.8 g (87% yield) of the expected product are obtained as an oil, following the procedure of Preparation IV, starting from 1.8 g (12.10 x 3 mol) of methyl 2-methoxypropanedioate and g (11.10-3 mol) of bis (1,1-dimethylethyl) - [[(6-aminohexyl) imino] methylene] bis-carbamate and chloroform are replaced by dichloromethane.

3 H NMR (CDCl 3): 1.35-1.65 (m, 26H), 3.2 (m, 2H), 3.35 (q, 2H), 3.45 (s, 3H), 3.8 ( s, 3H), 4.3 (s, 1H), 6.7 (t, 1H), 8.3 (t, 1H), 11.5 (s, 1H).

PREPARATIVE EXAMPLE IX

J 25 1- (1,1-Dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -13-methoxy-12-oxo-2,4,11-triazatetradec-2 * »• 'Enidioate * · • 4 g (89% yield) of the expected product are obtained as an oil following the procedure of Preparation III, starting from 4.6 g (9.4 * 103 moles) of the product of Preparation Example ·, * VIII. .

3 H NMR (CDCl 3): 1.3-1.65 (m, 26H), 3.1-3.5 (m, 4H), 3.6 (s, 1 H, 3H), 4.3 (s, 1H ), 6.7 (t, 1H), 8.4 (t, 1H), 11.5 (broad s, :: 35 h).

19, 112211

PREPARATIVE EXAMPLE X

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] -13-methoxy-12,14-dioxo- 2,4,11 # 15,20,24-Hexaza-pentacos-5 2-enidioate 4 g (63% yield) of the expected product are obtained as an oil following the procedure of Preparation IV and starting from 3.77 g (8.10). (3 moles) of the product obtained in Preparation Example 10 IX and 2.7 g (8- 10'3 moles) of 1,1-dimethylethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl] -2 , 6-diazadecanoate and chloroform are replaced by dichloromethane.

1 HNMR (CDCl 3): 1.3-1.8 (m, 50H), 3.0-3.45 (m, 12H), 3.6 (s, 15 3H), 4.1 (m, 1H), 4.8 and 5.3 (broad s, 1H), 6.9 (broad s, 2H), 8.3 (t, 1H), 11.5 (s, 1H).

Example 3 N- [4 - [(3- (Amino) propyl] amino] butyl] -N * - [6 - [(amino-20-iminomethyl) amino] hexyl] -2-methoxypropanediamide tris (trifluoroacetate)> · * 2 86 g (89% yield) of the expected product are obtained in the form of an oil,

following the procedure of Example 1 and starting; Is determined from 3.47 g (4.3 x 10 3 moles) in Preparative Example X.

product obtained.

1 HNMR (Dimethylsulfoxide-d 6): 1.25-1.70 (m, 12H), 1.9 (m, 2H), 2.85-3.2 (m, 12H), 3.25 (s, 3H) , 4.1 (s, 1H), 6.8-8.7 (m, 12H).

13 C NMR (CD 3 OD): 24.2; 25.3; 27.36; 27,37; 29.9; 30.2; 37.9; 39.2; 40.2; 42.5; 45.7; 58.6; 158.7; 169.4; 169.8.

* · # • »·» 1 1 20 112211

PREPARATIVE EXAMPLE XI

1- (1,1-dimethylethyl) -14-methyl-6 - [[(1,1-dimethylethoxy) carbonyl] -13-methoxy-12-oxo-2,6,11-triaza-tetradecanedioate 5 L, 5 g (18% yield) of the expected product are obtained as an oil, following the procedure of Preparation IV, starting from 2.62 g (17.7 x 10 3 mol) of methyl 2-methoxypropanedioate and 3.45 g. (10 -10'3 moles) 1,1-10 dimethylethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl] -2,6-diazadecanoate.

1 HNMR (CDCl 3): 1.4-1.8 (m, 24H), 3.1-3.4 (m, 8H), 3.5 (s, 3H), 3.8 (s, 3H), 4 , 3 (s, 1H), 4.8 and 5.3 (broad s, 1H), 6.7 (s, 1H).

15

PREPARATIVE EXAMPLE XII

1- (1,1-Dimethylethyl) -6 - [[(1,1-dimethylethoxy) carbonyl] -13-methoxy-12-oxo-2,6,11-triazatetradecanedioate 20 1.18 g (81% yield) ), the expected product is obtained in the form of an oil, * · following the procedure of Preparation Example III and starting from 1.5 g (3.16 · 10-3 mol) of the product obtained in Example XI.

3 H NMR (CDCl 3): 1.4-1.8 (m, 24H), 3.15-3.35 (m, 8H), 3.5 (s, 3H), 4.3 (s, 1H), 5.2-5.7 (ds, 1H), 6.3-6.9 (ds, 1H).

* * * »

PREPARATIVE EXAMPLE XIII

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carboxyl] amino] -22 - [(1,1-dimethylethoxy) carbonyl] -15-methoxy -14,16-dioxo-2,4,13,17,22,26-hexa-aza-heptacos-2-enidioate

I I I

1.4 g (65% yield) of the expected product are obtained as an oil 35 following the procedure of Preparative Example IV and starting from 1.18 g (2.56 · 10'3 moles) of the product of Preparation XII. .

21 112211 1 H NMR (CDCl 3): 1.3-1.8 (m, 54H), 3.1-3.7 (m, 15H), 4.1 (s, 1H), 4.8 and 5.3 ( broad s, 1H), 6.9 (broad s, 2H), 8.3 (t, 1H), 11.5 (s, 1H).

Example 4 N- [4 - [(3- (amino) propyl] amino] butyl] -2-methoxy-N '- [8- [(aminoiminomethyl) amino] octyl] propanediamide tris (trifluoroacetate) 0.89 g (70% yield) of the expected product is obtained in the form of an oil, following the procedure of Example 1, starting from 1.37 g (1.65-10. 3 moles) of the product of Preparation XIII.

1 HNMR (Dimethylsulfoxide-d 6): 1.2-1.7 (m, 16H), 1.9 (m, 15H), 2.9-3.10 (m, 12H), 3.3 (s, 3H). ), 4.1 (s, 1H), 6.8-8.7 (m, 12H).

CNMR (D 2 O): 23.8; 25.0; 26.5; 27.0; 27.3; 28.5; 29.1 (2C); 37.5; 39.8; 40.2; 42.5; 45.1; 48.2 58.4; 63.9; 82.5; 157.8; 169.8, 170.2.

PREPARATIVE EXAMPLE XIV

1- (1,1-Dimethylethyl) -15-methyl-6 - [(1,1-dimethylethoxy) carbonyl] -13-oxa-12-oxo-2,6,11-triazapentadecaa, 25 nidioate * ·

A solution of 7.4 g (21.4 x 10 3 mol) of 1,1-dimethyl-ethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl] -2,6-diazadecanoate ml of toluene is added with stirring to a solution of 4.5 g (21.4 x 10 ~ 3 moles) »methyl [(phenoxycarbonyl) oxy] acetate in 100 ml of toluene, and the reaction mixture is refluxed for 15 hours. The solvent is evaporated under reduced pressure and the residue obtained is purified by chromatography on 35 silica using methyl cyclohexane / ethyl acetate (7/3, v / v) as eluant followed by 22,112,111 ethyl acetate to give 8.4 g (85% yield). ) expected product as an oil.

1 HNMR (CDCl 3): 1.45-1.65 (m, 24H), 3.05-3.25 (m, 8H), 3.8 (s, 3H), 4.7 (s, 2H), 4 , 9 and 5.3 (broad s, 1H).

5

PREPARATIVE EXAMPLE XV

1- (1,1-Dimethylethyl) -6 - [(1,1-dimethylethoxy) carbonyl] -13-oxa-12-oxo-2,6,11-triazapentadecanedioate 6.7 g (81% yield) the expected product is obtained as an oil following the procedure of Preparation Example III starting from 8.45 g (18.3 * 10 3 moles) of the product obtained in Preparation Example XIV.

1 HNMR (CDCl 3): 1.3-1.8 (m, 24H), 3.10-3.25 (m, 8H), 4.7 (s, 15 2H), 5.0 (t, 1H), 6.8 (s, 1H).

PREPARATIVE EXAMPLE XVI

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -21 - [(1,1-dimethylethoxy) carbonyl] -14-20 oxa-12,15-dioxo -2,4,11,16,21,25-Hexaza-hexahexacos-2-enidioate * ·> · '3.85 g (73% yield) of the expected product are obtained in the form of an oil, following the procedure of Preparation IV * »25 and starting from 3 g (6.7 * 10'3 moles) of the preparation example * * f * j * · '. of product XV and 2.4 g (6.7 * 10 ~ 3 moles) * * bis (1,1-dimethylethyl [[(6-aminohexyl) imino] methyl * · · ethylene] biscarbamate.

3 H NMR (CDCl 3, D 3 O): 1.35-1.65 (m, 50H), 3.10-3.30 (m, 12H), 30 4.55 (s, 2H), 5.2-5.5 (broad s, 1H), 6.4 (broad s, 1H).

* 'I t »»' I I »i» 23 112211

PREPARATIVE EXAMPLE XVII

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -23 - [(1,1-dimethylethoxy) carbonyl] -16-oxa-14,17- dioxo-2,4,13,18,23,27-hexazazaoctacos-2-ene-5 dioate 1.56 g (57% yield) of the expected product is obtained as an oil following the procedure of Preparation IV and starting with 1.5 g. (3.35 * 10'3 moles) of the product obtained in Preparation Example 10, and 1.3 g (3.35 * 103 moles) of bis (1,1-dimethylethyl - [[(8-amino-octyl) ) imino] metylee-ni] biscarbamate.

1 H NMR (CDCl 3): 1.40-1.60 (m, 54H), 3.10-3.40 (m, 12H), 4.5 (s, 2H), 5.1-4.4 (broad s) , 1H), 6.3 (br s, 1H), 8.3 δ (t, 1H), 11.5 (br s, 1H).

Example 5 2 - [[[4 - [[3- (amino) propyl] amino] butyl] amino] carbonyl] oxy-N- [6 - [(aminoiminomethyl) amino] hexyl] acetamide 20 tris (trifluoroacetate) 2.61 g (72% yield) of the expected product are obtained as an oil; following the procedure of Example 1 and starting from 3.85 g (4.9 x 10'3 moles) of the product of Preparation XVI 25.

1 HNMR (Dimethylsulfoxide-d 6): 1.25-1, 55 (m, 12H), 1.9 1 · (m, 2H), 2.80-3.10 (m, 12H), 4.35 (s, 2H), 5.5-8.6 (m, 12H).

13 C NMR (D 2 O): 23.6; 24.5; 26.2; 26.3; 26.7; 28.6; 29.0; : 30 37.3; 39.8; 40.6; 41.9; 45.2; 48.2; 63.7; 157.5; 158.2; 171.4.

»*» · 24 112211

Example 6 2 - [[[4 - [[3- (Amino) propyl] amino] butyl] amino] carbonyloxy] -N- [8- [(aminoiminomethyl) amino] octyl] acetamide tris (trifluoroacetate) 50 96 g (66% yield) of the expected product are obtained as an oil, following the procedure of Example 1, starting from 1.56 g (1.9-103 mol) of the product of Preparation XVII.

1 HNMR (Dimethylsulfoxide-d 6): 1.2-1.6 (m, 16H), 1.9 (m, 2H), 2.8-3.2 (m, 12H), 4.35 (s, 2H) ), 6.8-8.6 (m, 12H).

13CNMR (D 6 O): 23.6; 24.5; 26.5; 26.6; 26.7; 28.6; 28.9 (2C); 29.0; 37.9; 40.0, - 40.6; 41.9; 45.2; 48.2; 64.7; 15 158.0; 159.0; 172.1.

PREPARATIVE EXAMPLE XVIII

1- (1,1-Dimethylethyl) -14-ethyl-3 - [[(1,1-dimethylethoxy) carbonyl] amino] -13-phenylmethoxy-12-oxo-20,4,11-triazatetradec-2 -enidioate 1.57 g (50% yield) of the expected product is obtained as an oil following the procedure of Preparative Example IV: starting from 1.3 g (5.46-10. 3 mol) of ethyl 2-phenylmethoxypropanedioate and 1.95 g (5.46.10-3 mol) of bis (1,1-dimethylethyl) - [[(6-aminohexyl) -1: 1, mino] methylene] biscarbamate.

1 HNMR (CDCl 3): 1.2-1.8 (m, 29H), 3.25 (q, 2H), 3.4 (q, 2H),. . 4.2 (q, 2H), 4.4 (s, 1H), 4.5-4.8 (dd, 2H), 6.7 (s, 1H), 7.3 (s, 5H), 8.3 (t, 1H), 11.5 (s, 1H).

»25 112211

PREPARATIVE EXAMPLE XIX

1- (1,1-Dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -13-phenylmethoxy-12-oxo-2,4,11-triazatet-Radek-2-enidioate 5 1.4 g (94% yield) of the expected product are obtained in the form of a yellowish oil following the procedure of Preparation Example III starting from 1.57 g (2.72 x 10 -3 mol) of the product obtained in Preparation XVIII.

1 H NMR (CDCl 3): 1.2-1.8 (m, 26H), 3.2-3.5 (m, 4H), 4.4 (s, 1H), 4.6-5.0 (dd) , 1H), 6.8 (s, 1H), 7.3 (s, 5H), 8.3 (t, 1H).

PREPARATIVE EXAMPLE XX

15 Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] -12,14-dioxo-13-phenylmethoxy -2,4,11,15,20,24-Hexaza-pentacos-2-enidioate 2.1 g (94% yield) of the expected product is obtained as an oil following the procedure of Preparation IV starting from 1.4 g ( 2.54 · 10-3 moles) of the product obtained in Preparation Example XIX and 0.88 g (2.54 · 10-3 moles) »1,1-dimethylethyl-10-amino-6 - [(1, 1-Dimethylethoxy) carbonyl] -2,6-diazadecanoate.

1 HNMR (CDCl 3): 1.3-1.8 (m, 50H), 3.1-3.5 (m, 12H), 4.3 (s, 1H), 4.8 (s, 2H), δ , 8-7.0 (m, 2H), 7.3 (s, 5H), 8.3 (t, 1H), δ, 5.5 (s, 1H).

30 PREPARATION EXAMPLE XXI

; * 'Bis (1,1-dimethylethyl) -3 - [((1,1-dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] -13-hyd- " oxy-12,14-dioxo-2,4,11,15,20,24-hexa-azapentacos-2-enidioate 35 * 0.1 g of 10% palladium on carbon catalyst is added to a solution of 1, 27 (1.45 x 10-3 moles) of Preparation Example 26 112211 Cat XX product in 120 mL ethanol The mixture is stirred at room temperature and under a hydrogen atmosphere for 2 hours at atmospheric pressure, then the catalyst is filtered off and the organic phase is evaporated to give 15 g. (88% yield) of an oily residue which is used without further purification to prepare the product of Example 7.

1 HNMR (CDCl 3): 1.2-1.8 (m, 50H), 3.1-3.6 (m, 12H), 4.3 (s, 1H), 4.6-5.3 (broad s) , 3H), 8.3 (s, 1H), 11.5 (s, 1H).

10

Example 7 N- [4 - [[3- (amino) propyl] amino] butyl] -2-hydroxy-N 1 - [6 - [(aminoiminomethyl) amino] hexyl] propanediamide tris (trifluoroacetate) 0.6 g ( 65% yield) of the expected product is obtained as an oil following the procedure of Example 1 starting from 1 g (1.27-10.3 mol) of the product of Preparation XXI.

1 HNMR (Dimethylsulfoxide-d 6): 1.2-1.6 (m, 12H), 1.85 (m, 2H), 2.7-3.2 (m, 12H), 4.3 (s, 1H) ), 6.8-8.6 (m, 12H).

13 C NMR (D 2 O): 23.5; 24.4; 26.0; 26.1; 26.2; 28.4; 28.8; 37.2; 39.1; 39.9; 41.7; 45.1; 48.0; 73.0; 154.8; 171.1; : 25,171.4.

, PREPARATIVE EXAMPLE XXII

1- (1,1-Dimethylethyl) -15-phenylmethyl-3 - [[(1,1-dimethyl, ethyl-ethoxy) carbonyl] amino] -12-oxo-2,4,11,14-tetra- Azapentadec-2-enidioate tt 1 · ·: A solution of 1.6 g (14 · 10 · 3 moles) of isobutyl chloroformate in 5 ml of tetrahydrofuran is added dropwise. to a solution cooled to -30 ° C and containing 35 g of carbobenzyloxyglycine 3 g (14 · 10-3 moles) and 2.8 g · 28 · 10 ~ 3 moles of N- methyl morpholine in 50 ml of tetrahydrofuran. The reaction mixture is stirred for 0.5 hour and a solution of 5.4 g (14.10-10 mol) of bis (1,1-dimethylethyl - [[(6-aminohexyl)) is added. imino] methylene] bis-carbamate in 20 ml of tetrahydrofuran Stirring is continued for 2 hours at -30 [deg.] C., then for 24 hours at 5 [deg.] C. The reaction mixture is filtered, then the filtrate is evaporated under reduced pressure and the resulting residue is purified by silica ethyl acetate / methylcyclohexane (1/1, v / v), 7.16 g (91% yield) of the expected product are obtained in the form of an oil.

1 HNMR (CDCl 3): 1.3-1.7 (m, 26H), 3.2 (q, 2H), 3.4 (q, 2H), 3.8 (d, 2H), 5.15 (s , 2H), 5.5 (broad s, 1H), 6.0 (broad s, 1H), 7.3 (s, 5H).

15 PREPARATIVE EXAMPLE XXIII

1,1-Dimethyl-ethyl-13-amino-3 - [[(1,1-dimethyl-toxicoxy) carbonyl] -amino] -12-oxo-2,4,11-triazatridec-2-enoate 3 g (98% yield) of the expected product are obtained as an oil following the procedure of Preparation Example XXI and starting from 7.1 g (13 · 10-3 moles) of the product obtained in Preparation XXII.

1 HNMR (CDCl 3): 1.3-1.6 (m, 28H), 3.25-3.45 (m, 6H), 7.3 (s, 25 1H), 8.3 (t, 1H), 11.5 (s, 1H).

*

,; 'PREPARATIVE EXAMPLE XXIV

* 'Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -21 - [(1,1-dimethylethoxy) carbonyl] -12,15-30 dioxo-2, 4,11,14,16,21,25-hepta-aza-hexahexacos-2-enidio, acetate (· · · - 4.3 g (13 · 10-3 moles) of bis (4-nitrotenyl) carbonate) is added in small portions to a solution of 5.3 g / · 35 (12 · 10 103 mol) of the product obtained in Preparation XXIII in 50 ml of anhydrous tetrahydrofuran. The reaction mixture is stirred for 1 hour at room temperature and a solution of 28 , 5 g (13.10 -3 mol) of 1,1-dimethylethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl] -2,6-diazadecanoate in 50 ml of anhydrous tetrahydrofuran are added dropwise Stirring is continued for 24 hours at room temperature and the solvent is evaporated under reduced pressure to give a residue which is purified by chromatography on silica using ethyl acetate as eluant to give 6.01 g (64% yield) of the expected product as an oil.

1 HNMR (CDCl 3): 1.3-1.7 (m, 50H), 3.1-3.35 (m, 12H), 3.8 (d, 2H), 4.8 and 5.8 (broad) s, 3H), 6.9 (t, 1H), 8.3 (t, 1H), 11.5 (s, 1H).

Example 8 N- [4 - [[3- (Amino) propyl] amino] butyl] -N '- [[[[6 - [(amino-iminomethyl) amino] hexyl] amino] carbonyl] methyl] uranium tris (trifluoroacetate) 4.75 g (86% yield) of the expected product are obtained as an oil following the procedure of Example 1 and starting from 6 g (7.6.10-3 mol) of the product of Preparation XXIV.

1 HNMR (Dimethylsulfoxide-d 6): 1.2-1.65 (m, 12H), 1.9 (m, 2H), 2.9-3.15 (m, 12H), 3.6 (d, 2H) ), 6.1 (t, 1H), 6.3 (t, 1H), 6.8-9 (m, 11H).

C 25 V 13 CNMR (dimethylsulfoxide-ds): 22.8; 23.7; 25.6; 25.8; 27.0; 28.3; 28.9; 36.1; 38.3; 38.5; 40.5; 42.7; 43.7; 46.5; 156.7; 157.9; 169.6.

I i

30 PREPARATION EXAMPLE XXV

1- (1,1-dimethyl-ethyl) -17-phenylmethyl-3 - [[(1,1-dimethyl-1,1-ethyl-ethoxy) -carbonyl] -amino] -14-oxo-2,4,13,16 -tetra-at- ", saheptadec-2-enidioate, Γ 35 2.33 g (74% yield) of the expected product are obtained as an oil following the procedure of Preparation Example IV starting from 1.35 g (6.47.10"). 3 moles) of carbobenzyl-29 112211 oxyglycine and 2 g (5.18-10.10 moles) of bis (1,1-dimethylethyl) - [[(8-amino-octyl) imino] methylene] bis-carboxylic acid. dichromate.

1 HNMR (CDCl 3): 1.25-1.70 (m, 30H), 3.25 (q, 2H), 3.4 (q, 2H), 3.8 (d, 2H), 5.15 ( s, 2H), 5.5 (broad s, 1H), 6.0 (broad s, 1H), 7.3 (s, 5H), 8.3 (t, 1H), 11.5 (s, 1H) ).

PREPARATIVE EXAMPLE XXVI

1,1-Dimethylethyl-15-amino-3 - [[(1,1-dimethylethoxy) -10 carbonyl] amino] -14-oxo-2,4,13-triazapentadec-2-enoate 2.16 g ( yield 100%) of the expected product is obtained as an oil following the procedure of Preparation Example XXI and starting from 2.33 g (4.04 x 10 3 mol) of the product obtained in Preparation Example XXV.

1 HNMR (CDCl 3): 1.3-1.8 (m, 32H), 3.25-3.45 (m, 6H), 7.3 (s, 1H), 8.3 (t, 1H), 11 , 5 (s, 1H). 1 2 3 4 5 6> 1 »

PREPARATIVE EXAMPLE XXVII

2

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -23 - [(1,1-dimethylethoxy) carbonyl] -14,17-4'-dioxo -2,4,13,16,18,23,27-hepta-azaoctacos-2-enidioate-5 *: ti 6 * ·. 1.5 g (45% yield) of the expected product are obtained as an oil, following the procedure of Preparation Example XXIV and starting from 1.79 g (4.04-103 mol) of the product obtained in Preparation Example XXVI.

Λ 30 H NMR (CDCl 3): 1.3-1.7 (m, 54H), 3.1-3.4 (m, 12H), 3.8 (d, *. * 2H), 4.8. 5.2 and 5.7 (broad s, 2H), 6.0 and 6.7 (broad s, 1H), 8.3 (t, 1H), 11.5 (s, 1H).

• i 3Θ 112211

Example 9 N- [4 - [[3- (Amino) propyl] amino] butyl] -N '- [[[[8- [(aminoiminomethyl) amino] octyl] amino] carbonyl] methyl] urea tris (trifluoroacetate) 1.14 g (82% yield) of the expected product are obtained as an oil, following the procedure of Example 1, starting from 1.5 g (1.84 x 10'3 moles) of the product of Preparation XXVII.

1 H NMR (Dimethylsulfoxide-d 6): 1.25-1.55 (m, 16H), 1.9 (m, 2H), 2.9-3.10 (m, 12H), 3.6 (s, 2H) ), 6.0-6.3 (broad s, 2H), 6.8-8.6 (m, 11H).

13CNMR (dimethylsulfoxide di): 22.9; 23.8; 26.0; 26.3; 15 27.1; 28.4; 28.5; 28.6; 29.1; 36.2; 38.4; 38.7; 40.7; 42.7; 43.8; 46.6; 156.7; 158.0; 169.6.

PREPARATIVE EXAMPLE XXVIII

1- (1,1-Dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino-12-oxo-2,4,11-triazatridec-2-enidioate

A solution of 0.67 g (5 x 10'3 moles) of ethyl oxalate acid chloride in 5 mL of dichloromethane is added dropwise to a solution of 1.6 g (4.5 x 10'3 moles) of bis (1, 1-25 dimethylethyl) - [[(6-aminohexyl) imino] methylene] bis-. carbamate and 0.6 g (6.10-3 mol) of triethylamine in 10.1 ml of anhydrous dichloromethane. The mixture is stirred for 1 * · hour at room temperature, then the solvent evaporates. The residue obtained is chromatographed under reduced pressure and the resulting residue is chromatographed on silica using hexane / ethyl acetate (2/1, v / v) as eluant to give 1.68 g (82% yield) of the expected product as an oil.

·. 1 HNMR (CDCl 3): 1.3-1.7 (m, 29H), 3.3-3.4 (m, 4H), 4.4 (q, 2H), 7.1 (t, 1H), δ , 3 (t, 1H), 11.5 (s, 1H).

: Γ 35 31 112211

PREPARATIVE EXAMPLE XXIX

1- (1,1-Dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -12-oxo-2,4,11-triazatridec-2-enidioate δ 1.2 g (yield) 75%) of the expected product is obtained in the form of an oil following the procedure of Preparation Example III and starting from 1.67 g (3.7 x 10 -3 mol) of the product obtained in Preparation Example XXVIII.

1 HNMR (CDCl 3): 1.25-1.7 (m, 26H), 3.3-3.5 (m, 4H), 7.4 (t, 1H), 8.5 (t, 1H), 11.5 (broad s, 1H).

PREPARATION EXAMPLE XXX

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -19 - [(1,1-dimethylethoxy) carbonyl] -12,13-15 dioxo- 2,4,11,14,19,23-Hexazazetetracos-2-enidioate 1.2 g (53% yield) of the expected product are obtained as an oil following the procedure of Preparation XXII starting from 1.4 g (3.5 · 103 moles) of the product obtained in Preparation XXIX and 1.05 g (3.10'3 moles) of 1,1-dimethylethyl-10-amino-6 - [(1,1-dimethyl-ethoxy) carbonyl]. -2.6-diatsadekanoaattia.

3 (CDCl 3): 1.3-1.7 (m, 50H), 3.1-3.4 (m, 12H), 4.8 and 5.3 (broad S, 1H), 7.5 (m, 2H). ), 8.3 (t, 1H), 11.5 (s, 1H).

25 *. ·. Example 10! N- [4 - [[3- (amino) propyl] amino] butyl] -N '- [6 - [(aminoimin * * * nomethyl) amino] hexyl] ethanediamide tris (trifluoroacetate) •: V 1.05 g (95% yield) of the expected product is obtained in the form of an oil; following the procedure of Example 1, starting from 1.2 g (1.6 · 10-3 moles) of the product obtained in Preparation Example * · · · XXX.

· 35 H NMR (dimethylsulfoxide-d5): 1.3-1.65 (m, 12H), 1.9 (m, 2H), 2.8-3.3 (m, 12H), 6.8-9 , 9 (m, 12H).

32 112211 13 CNMR (dimethylsulfoxide-d 6, D 2 O): 23.9; 24.7; 26.3; 26.4; 26.5; 28.7; 29.0; 37.5; 39.7; 40.4; 42.0; 45.4; 48.1; 157.7; 161.8; 162.1.

5 PREPARATION EXAMPLE XXXI

1- (1,1-Dimethylethyl) -13-ethyl-6 - [(1,1-dimethylethoxy) carbonyl] -12-oxo-2,6,11-triazatridecanedioate 2.31 g (89% yield) of the expected product are obtained. as an oil, following the procedure of Example XXVIII and starting from 2 g (5.8 x 10 3 mol) of 1,1-dimethylethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl] -2, 6-diaza-decanoate and 1.03 g (7.54-103 moles) of ethyl oxalate acid chloride.

1 HNMR (CDCl 3) δ 1.25-1.65 (m, 27H), 3.1-3.35 (m, 8H), 4.35 (q, 2H), 4.75 and 5.25 (broad S, 1H), 7.2 (s, 1H).

PREPARATIVE EXAMPLE XXXII

1- (1,1-Dimethylethyl) -6 - [(1,1-dimethylethoxy) carbonyl-20] -12-oxo-2,6,11-triazatridecanedioate 2.16 g (100% yield) of the expected product are obtained as an oil , ·, · Following the procedure of Example III and starting from 2.31 g (5.19 · 10 ~ 3 moles) of the product obtained in Preparation Example ·, 25 XXXI.

; 1 H NMR (CDCl 3): 1.25-1.70 (m, 24H), 3.1-3.4 (m, 8H), 4.8-5.25 (ds, 1H), 7.5 (broad s, lH).

• <·

PREPARATIVE EXAMPLE XXXIII

: · 30 Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] 21 - [(1,1-dimethylethoxy) carbonyl] -14,15-, ·. dioxo-2,4,13,16,21,25-hexahaza-hexahexacos-2-enidioate> · * · '· * 0.9 g (48% yield) of the expected product are obtained as an oil 35 following the procedure of Example IV and starting from 1 g (2.4 · 10'3 moles) of the product of Preparation XXXII and 0.92 g (2.4 * 10'3 moles) of 33,112,211 bis (1.1). dimethylethyl) - [[(8-aminooctyl) imino] methylene-phenylene] biscarbamate.

1 HNMR (CDCl 3): 1.3 (t, 3H), 1.4-1.7 (m, 24H), 3.0-3.4 (m, 8H), 4.25 (q, 2H), 4 , 4 (s, 1H), 4.5-4.7 (2d, 2H), 5.0 (s, 5H), 6.7 (S, 1H), 7.3 (m, 5H).

Example 11 N- [4 - [[3- (Amino) propyl] amino] butyl]-] - [8- [(aminoiminomethyl) amino] octyl] ethanediamide tris (trifluoroacetate) 0.15 g ( 18%) of the expected product is obtained as an oil following the procedure of Example 1 starting from 0.9 g (1.15 * 10 3 moles) of the product of Preparation XXXIII.

1 H NMR (dimethylsulfoxide-d 6): 1.25-1.50 (m, 16H), 1.9 (m, 2H), 2.9-3.15 (m, 12H), 6.8-8.8 ( m, 12 H).

X 3 CNNR (D 2 O): 23.8; 24.5; 26.1; 26.5; 26.7; 28.6; 28.92; 20.95; 29.0; 37.3; 39.5; 40.4; 42.0; 45.2; 48.1.

*

V. PREPARATION EXAMPLE XXXIV

1- (1,1-Dimethylethyl) -14-ethyl-6 - [(1,1-dimethylethoxy) carbonyl] -12-oxo-13-phenylmethoxy-2,6,11-triaza-25-tetradecanedioate

The expected product is obtained in 35% yield after chromatography on silica (eluent: .., methyl cyclohexane / ethyl acetate, 8/2), followed by the procedure of Preparation IV and starting with 3 g. of ethyl 2- (phenylmethoxy) propanedioate (12.6 x 10 -3 moles) and of 1,1-dimethylethyl-10-amino-6- (4.4 g, 12.6 x 10 3 moles). [(1,1-Dimethylethoxy) carbonyl] -2,6-diazadecanoate.

1 H NMR (CDCl 3): 1.3 (t, 3H); 1.4-1.7 (m, 24H), 3.0-3.4 (m, · *: 8H), 4.25 (q, 2H), 4.4 (s, 1H), 4.5 -4.7 (2d, 2H), 5.0 (s, 1H), 6.7 (s, 1H), 7.3 (m, 5H).

34, 112211

PREPARATION EXAMPLE XXXV

1- (1,1-Dimethylethyl) -6 - [(1,1-dimethylethoxy) carbonyl] -12-oxo-13-phenylmethoxy-2,6,11-triazatetradecanedioate 2.46 g (4, 3 x 103 moles) of the product of Preparation XXXIV is dissolved in ethanol / water (1/1, v / v) and 258 mg of sodium hydroxide is added. The reaction mixture is stirred at room temperature for 24 hours, after which 10 ml of water and 20 ml of chloroform are added and the mixture is acidified with 1N hydrochloric acid to pH 2. It is extracted with chloroform and the organic phase is dried over magnesium sulphate and concentrated under reduced pressure. product, yield 76%.

1 HNMR (CDCl 3): 1.4-1.7 (m, 24H), 3.0-3.3 (m, 8H), 4.4-4.5 (m, 1H), 4.7-5 , 1 (2d, 2H), 5.0 (s, 1H), 6.65 (s, 1H), 7.4 (m, 5H).

PREPARATION EXAMPLE XXXVI

20 Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -22 - [(1,1-dimethylethoxy) carbonyl] -1,14,16-dioxo-15- phenylmethoxy-2,4,13,17,22,26-hexahaza-azaheptacos-2-enidioate 1 2 3 4 5 6 7 8 9 10 11

· «I

The expected product is obtained as an oil in 59% yield after 2 * 'purification by chromatography on silica 3! (eluent: methylcyclohexane / ethyl acetate mixture, 4/7/3), follow the procedure of Preparation Example IV, starting from 1.67 g (3.11-10.3 mol) of the product of Preparation 6 Example XXXV and 1.20 of g (3.11-10.10 moles) of 7 · bis ((1,1-dimethylethyl) - [[(8-amino-octyl) imino] methylene] bis-carbamate.

9 H NMR (CDCl3): 1.2-1.8 (m, 54H), 3.0-3.5 (m, 12H), 4.3 (s, 10H), 4.8 (m, 2H) , 5.1 (s, 1H), 6.9-7.1 (m, 2H), 7.4 (m, 5H), 11 8.3 (t, 1H), 11.5 (s, 1H) .

35, 112211

PREPARATION EXAMPLE XXXVII

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -22 - [(1,1-dimethylethoxy) carbonyl] -14,16-dioxo-15-hydroxy- 2,4,13,17,22,26-Hexa-aza-heptacos-2-5 enidioate

The expected product is obtained in 55% yield following the procedure of Preparation Example XXI and starting from 1.64 g of the product obtained in Preparation XXXVI.

1 HNMR (CDCl 3); 1.3-1.7 (m, 54H), 3.0-3.4 (m, 8H), 4.4 (s, 1H), 5.0 (s, 1H), 7.2 (s, 1H), 7.35 (s, 1H), 8.3 (t, 1H), II, 5 (s, 1H).

Example 12 N- [4 - [[3- (amino) propyl] amino] butyl] -2-hydroxy-N '- [8- [(aminoiminomethyl) amino] octyl] propanediamide tris (trifluoroacetate)

The expected product is obtained as an oil in 67% yield after purification by MPLC using RPIS-grafted silica (eluent: water / acetonitrile / trifluoroacetic acid mixture, 7.5 / 1.5 / 1) The procedure of Example 1 and starting from 0.81 g of the product of Preparation XXXVII.

1 H NMR (DMSO-d 6): 1.1-1.7 (m, 16H), 1.9 (m, 2H), 2.8-3.2 (m, 12H), 4.3 (s, 1H), 6.9-8.7 (m, 12H).

CNMR (DMSO-d 6): 14.8; 20.9; 22.2; 23.1; 25.3; 25.9; 28.5; 28.7; 28.9; 36.8; 38.1; 43.8; 46.2; 60.1; 71.8; 30,157.3; 166.2; 166.5.

· »» T »36 112211

PREPARATION EXAMPLE XXXVIII

1- (1,1-Dimethylethyl) -14-ethyl-6 - [(1,1-dimethylethoxy) carbonyl] -13-fluoro-12-oxo-2,6,11-triazatetradecanedioate

Prepare a solution of 1.7 g (11.10-3 moles) of ethyl 2-fluoropropanedioate and 2.22 g (22.103 moles) of N-methylmorpholine in 50 ml of anhydrous tetrahydrofuran (THF) and 5 ml. in anhydrous dimethylformamide. The mixture is then cooled to -20 ° C and a solution of 1.6 ml (12.10 x 3 mol) of isobutyl chloroformate in 5 ml of THF is added. The resulting mixture is stirred for 30 minutes at -20 ° C. Followed by the addition of a solution of 3.9 g (11.10-3 mol) of 1,1-dimethyl-15-ethyl-10-amino-6 - [(1,1-dimethylethoxy) carbonyl]. - 2,6-Diazadecanoate in 30 ml of THF Stirring is continued for 2 hours at -20 ° C and then for 12 hours at room temperature The reaction mixture is filtered and the filtrate is concentrated under reduced pressure The residue obtained is purified by chromatography on silica (eluent). : methyl cyclohexane / ethyl acetate, 7/3), the expected product is obtained in the form of an oil in 34% yield.

1 H NMR (CDCl 3): 1.34 (t, 3H), 1.4-1.7 (m, 24H), 3.0-3.4 (m, 8H), 4.3 (m, 2H), 5.0 (s, 1H), 5.25 (d, 1H), 6.55 (s, 1H).

*: 25

. ·. ·! PREPARATION EXAMPLE XXXIX

1- (1,1-Dimethyl-ethyl) -6 - [(1,1-dimethylethoxy) carbonyl] -13-fluoro-12-oxo-2,6,11-triazatetradecanedioate: Prepare a solution of 1, 8 g (3.8 x 10 ~ 3 moles) of the product obtained in Preparation Example XXXVIII in 6 ml of aqueous 1N sodium hydroxide and 20 ml of dimethoxyethane. Stir at room temperature for 1 hour, then add 10 ml of water and 20 ml of dichloromethane; 35 dan and the mixture acidified with 1N hydrochloric acid to pH · 1. 2. It is extracted twice with 20 ml of dichloromethane and the organic phase is dried over magnesium sulfate and concentrated under reduced pressure to give 1.55 g of the expected product as an oil (yield). 91%).

1 HNMR (CDCl 3): 1.4-1.7 (m, 24H), 3.0-3.35 (m, 8H), 5.0 (s, 1H), 5.3 (d, 1H), δ , 5 (s, 1H).

5

PREPARATION EXAMPLE XL

Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] -12,14-dioxo-13-fluoro- 2,4,11,15,20,24-Hexazapenta-10-cos-3-enidioate

A solution of 0.29 mL (2.2 x 10'3 mol) of isobutyl chloroformate in 5 mL of THF is added dropwise to a solution cooled to -20 ° C with 1 g of 15 (2). , 2 x 10'3 moles) of the product of Preparation XXXIX and 4.5 g (4.4 x 10 3 moles) of N-methylmorpholine in 30 ml of anhydrous THF. The mixture is stirred for 30 minutes at -20 ° C: followed by addition of a solution of 0.876 g (2.2 x 10 ~ 3 moles) of bis (1,1-dimethylethyl) - [[(6-20 aminohexyl) imino] methylene] biscarbamate and 0.34 ml (2 , 2 · 10'3 moles) of triethylamine in 5 ml of THF The temperature is maintained at -20 ° C for 2 hours, then the mixture is brought to room temperature and stirring is continued for 12 hours. The residue is purified by chromatography on silica (eluent: ethyl acetate / cyclohexane 7/3) to give 1.43 g of the expected product as an oil (yield 81). %).

1 H NMR (CDCl 3): 1.3-1.7 (m, 50H), 3.0-3.45 (m, 12H), 5.0 (s, 30H), 5.2 (d, 1H), 6.8-7.1 (m, 2H), 8.3 (t, 1H), 11.5 (s, 1H). 1: I »i · 38 112211

Example 13 N- [4 - [[3- (amino) propyl] amino] butyl] -N '- [6 - [(aminoiminomethyl) amino] hexyl] -2-fluoro-propanediamide tris (trifluoroacetate) δ 263 mg expected the product is obtained in the form of an oil (71% yield) after purification by MPLC using RP18-grafted silica (eluent: water / acetonitrile / trifluoroacetate mixture 7.5 / 2 / 0.5) following the procedure of Example 10 and starting from 0.4 g (0.5 x 10 ~ 3 moles) of the product obtained in Preparation XL.

1 HNMR (DMSO-d 6): 1.25-1.55 (m, 12H), 1.9 (m, 2H), 2.7-3.15 (m, 12H), 5.2 (d, 1H) , 6.9-8.7 (m, 12H).

15 ”CNMR (D 2 O / Dioxane-d 8): 23.65? 24.53; 26.17; 26.25; 28.55; 28.79; 37.34; 39.32; 40.08; 41.86; 45.22; 48.09; 87.00; 89.66; 157.0; 166.2; 166.7.

PREPARATIVE EXAMPLE XLI

20 Bis (1,1-dimethylethyl) -3 - [[(1,1-dimethylethoxy) carbonyl] amino] -22 - [(1,1-dimethylethoxy) carbonyl] -15-! Fluoro-14,16-dioxo-2,4,13,17,22,26-hexazazaheptacos-2-phenylenedioate 0.513 g of the expected product is obtained in the form of an oil (61% yield), *, after purification chromatographed on silica (eluent: ethyl acetate / cyclohexane mixture, '6/4'), follow the procedure of Preparation Example XXII and start from 0.460 g (1.02 x 10 ° mole) in the preparation of 30 x XXXIX of the product obtained and 0.395 g (1.02 x 10'3 moles) of bis (1,1-dimethylethyl) - [[(8-amino-octyl) imino] methylene] bis-carbamate.

1 HNMR (CDCl 3) δ 1.2-1.7 (m, 54H), 3.0-3.4 (m, 12H), 5.0x1.5 (s, 1H), 5.2 (d , 1H), 6.8-7.1 (m, 2H), 8.3 (t, 1H), 11.5 (s, 1H).

1 · * ·

• · I

39, 112211

Example 14 N- [4 - [[3- (Amino) propyl] amino] butyl] -2-fluoro-N 1 - [8- [(aminoiminomethyl) amino] octyl] propanediamide Tris (Trifluoroacetate)

The expected product is obtained as an oil after purification by MPLC using? P18-grafted silica followed by the procedure of Preparation Example XXII and starting from 0.460 g (1.02110 3 moles) of the product obtained in Preparation 10 XLI. .

1 HNMR (DMSO-d 6): 1.25-1.50 (m, 1H), 1.84 (m, 2H), 2.7-3.15 (m, 12H), 5.20 (d, 1H) , 6.9-8.7 (m, 12H).

13CNMR (D 2 O / dioxane-d 8): 23.66; 24.55; 26.18; 26.28; 15 26.57; 27.48; 28.86; 37.35; 39.31; 40.21; 40.33; 45.23; 48.10; 87.26 to 89.86; 157.0; 166.2; 166.7.

Example 15 N- [4 - [[3- (Amino) propyl] amino] butyl] -N '- [6 - [(aminoimino-20-methyl) amino] hexyl] propanediamide tris (hydrochloride): Method A:; 1 g (1.41103 mol) of the product obtained in Example 1 is dissolved in a mixture of 7 ml of 10 M hydrochloric acid and 50 ml of distilled water, and the resulting solution is then lyophilized. This step is repeated twice. The lyophilized residue is taken up in a mixture of 9 ml of ethanol and 1 ml of methanol. The resulting crystals are filtered after 24 hours at room temperature, rinsed with isopropyl ether and dried in vacuo to give the expected; product as white crystals, yield 68%. Mp 130 ° C.

Method B: '... 2.5 g (3.28110'3 moles) of the product obtained in Preparation Example IV are dissolved in 25 ml of methanol saturated with 40-12211 ml of hydrogen chloride. After stirring overnight at room temperature, the solution is concentrated under reduced pressure and the residue is redissolved in 10 ml of water, followed by lyophilization. The resulting compound 5 is recrystallized from ethanol / methanol (9/1) to give a crystalline product, yield 52%.

Example 16 2 - [[[4 - [[3- (Amino) propyl] amino] butyl] amino] carbonyl-10-yloxy] -N- [6 - [(aminoiminomethyl) amino] hexyl] acetamide tris (hydrochloride)

The expected product is obtained in the form of crystals, yield 58%, following the procedure of Example 15 (Method B) 15 and starting from the product of Preparation XVI. Mp 148 ° C.

The immunosuppressive activity of the products obtained according to the invention was determined using the graft-versus-host reaction. B6D2Fl-male. mice (C57B1 / 6 x DBA / 2 first generation hybrids) are immunosuppressed by intraperitoneal (i.p.) injection of cyclophosphamide. After three days (experiment, day 0: DO), they are intravenously dosed with 4x107 * 25 C57Bl / 6 mouse spleen cells. The animals are then divided into groups of at least 8 animals and treated daily from day 1 (D1) to day 5 (D5) and day 7 (D7) to day 10 (D10) ·, *. i .p. Only vehicle is administered to the control group. Mortality is monitored until the 60th (D60). The results, expressed as the average lifetime days at the dose in question, are summarized in Table I, where the results given are significant by the Logrank test (probability £ 5%).

. '!! By way of comparison, Table I also shows the values obtained with known products: 15-deoxyspergualin (DSG), cyclosporin A, commonly used in comparative immunosuppressant therapy, and the product of Example 1 of EP-A-0 105 193. This comparison shows that the products made according to the invention are up to 250 times more active than those known in the art. In particular, the product of Example 1 of the invention is significantly active as early as 0.1 mg / kg, while the reference compound of Example 1 of EP-A-0 105 195 is significantly active only as of 1 mg / kg, deoxyspergualine starting at 0.3 mg / kg and cyclosporine 10 A starting at 25 mg / kg.

In addition, the compounds prepared according to the invention have significantly higher solution stability than those known in the art, especially 15-deoxyspergua-15.

The products of the invention are useful as therapeutics as healing or preventive immunosuppressants, in particular for preventing allogeneic or xenogeneic organ rejection or graft-versus-host blood-donation or non-graft-versus-host reactions. ,: in the treatment of genetically determined or acquired autoimmune diseases or chronic inflammatory diseases, as well as in any pathological condition in which an immune disorder, which is the cause or factor for the deterioration of the clinical condition.

The products of the invention may also be administered in combination with cytotoxic anticancer drugs: to limit their side effects and to reduce the occurrence of protective antibodies produced by the patient in connection with the administration of products of biotechnological origin, in particular recombinant cytokinins or monoclonal antibodies and polyclonal antibodies. .

42, 112211

The products of the invention can be used as curative agents in the treatment of parasitic diseases, especially malaria.

The products of the invention may be administered orally, by injection, in particular by intramuscular or intravenous injection, topically, in particular as a topical ointment or as eye drops, transdermally, suppository or by inhalation.

TABLE I

activity

Example "n A Dose (mg / kg) Lifetime (day) 16 CH, Θ.1 27.6 Θ.3 48.5 2 8 CH, 3 47.6 3 6 CH (OCH 3) 3 55.3 4 8 CH (OCH 3) 3 27, 6 5 6 CH, 0 3 57.9 7 6 CH (OH) 3 53.Θ 8 6 CH, NH 0.3 39.8 16 6 single-sided 1 52.9 11 8 single-sided 3 59.4 12 8 CH (OH) 1 55.6 13 6 CHF 3 66.0 15-deoxyspergualine 1 43.1

Cyclosporin A 25 36.6 · 'ΕΡ-Α-Θ 105 193 e.g. 1 1 32.0 * · 15-deoxyspergual11ni 0.3 32, i # __________ • 111 1 "~' ''" 11 ~~ 1 The compounds of the invention are in the tris (trifluoroacetate) form.

· »'· · * ·

I | I

· 1 * ♦ * · * · I t i • t t »· I» ♦ •

Claims (4)

A process for the preparation of a therapeutically applicable compound belonging to the genus of the 15-deoxypergualin analogues having the formula I nh o o J / <CHZ> n. II 11 (CH2) 4. ^ (CH2) 3x H n N ^ NH2 2. I I I 10. HHH (I) wherein n is 6 or 8 and A is a bond, CH 2, CH (OH) -, CHF, CH (OCH 3), CH 2 NH or CH 2 O group, or an acid addition salt thereof, characterized hence, in step (i) of Process Alternative A, a compound of formula II is reacted with R II 1N NHRAAh / (CK ^Nb „» I I n n n is 6 or 8 and R3 is an amino protecting group, with a (Iii) wherein X is a chlorine atom or the group OH, A is a simple bond, CH2, CHF, CH (OCH2C6H5) or CH (OCH 3) group and R 2 is a linear or branched C 1 -C 3 alkyl group or a phenylmethyl group, in an organic solvent in the presence of the carboxy group activator and a nucleophilic agent at a temperature of 0 ° C. 40 ° C, when the molar ratio of the compounds is about 1 mole of a compound of formula II / 1 mole of a compound of formula III, to obtain a compound of formula IV 1 (1) '' (CH2 AA2 R2 (IV)) where R1 # r2 and n are the same as above and A is a simple bond, CH2, CHF, CH (OCH2C6H5) or CH (OCH3), (ii) the compound formed with Formula IV is saponified in an organic solvent in the presence of a strong base to obtain a compound of formula V R 1 NH 1 0 Anh / (CH 2 NH 2 A 2 OH (V) where R x and n denote the same as above and A is a simple bond, CH 2, CHF, CH (OCH 2 C 6 H 5) or CH (OCH 3), (iii) the formed compound of formula V is condensed with an amine of formula VI 20 (CH 2) 4 r 1 (VI): R 1 'wherein R 3 is the same as above, under conditions. * which are the same as above in step (i), to obtain | a compound of formula VII ..: r,:: -NH oo R. -h ^ nh / (CH2> 3 ^ nh / R 'r, (vii) 1' where R3 and n are the same as above and A is single bond, CH2, CHF, CH (OCH2CsH5) or CH (OCH3), (iv) from a compound of formula VII wherein A is the group -t CH (OCH2C6H5) - is removed as needed protecting by catalytic hydrogenation to obtain a compound of formula VII, wherein A is a CH (OH) group, (v) from a compound of formula VII obtained in step (iii) or (iv) and where A is a simple bond, CH 2, CHF, CH (OH) or CH (OCH 3), the protecting group R 3 is removed by a reaction with a strong acid, to obtain an addition salt of a compound of formula I, where A bond, CH 2, CHF, CH (OH) or CH (OCH 3), and (vi) said compound of Formula I is obtained, as needed, in the form of a free base by a reaction with a strong base and then other addition salts are obtained from said free bass, or In step (i) of process alternative B, a compound of formula VI is reacted with (CH 2) a 2 (CH 2) 3 (VI) h 2 n 2 -nh * 1 wherein R 3 is a protecting group as mentioned above, with an acid or acid chloride of formula IIIo: (m):: 25 x! where X is a chlorine atom or the group OH, A is a simple bond, CH 2, CHF, CH (OCH 2 C 6 H 5) or CH (OCH 3) group j: and R 2 is a linear or branched C , in an organic solvent in the presence of the carboxy group activator and a nucleophilic agent at a temperature of 0 ° C - 40 ° C, when the mole ratio of the compounds is 1 mole of a compound of formula * VI / 1 mole of a compound of the formula III, to obtain a compound of the formula VIII (CH2) 4 \ R <CH2> 3 \ ^ R1 R 2 and R 2 are the same as above and A is a simple bond, CH 2, CHF, CH (OCH 3) or CH (OCH 2 C 6 H 5) group, (ii) the compound of formula VIII is saponified in an organic solvent in the presence of a strong base to obtain a compound of formula IX and high (CH 2, a Wherein R 2 is the same as above and A is a simple bond, CH 2, CHF, CH (OCH 3) or CH (OCH 2 C 6 H 5), (iii) the compound of formula IX formed is condensed with an amine of the formula II '' R: 1 'n R' (CH) 25 '- n' nh '* n' (II | 2 t 'where n is 6 or 8 and R3 represents the same as above, under conditions which are the same as above in step 30 (i), to obtain a compound of formula VII], R 1: NH 0 O: R'.A ™ / (CH 2> n \ AA. __ ^ (CH2) 3 \ - ^ R1 N NH ^ n ^ NH NH ^ 'N' 'NH; *' L (VII) 35, ·· *, where R3 and n are the same as above and A is a simple (iv) from a compound of formula VII wherein A is a CH (O-CH 2 C 6 H 5) group, if necessary, the protection is removed by catalytic hydrogenation to obtain a compound of formula VII, wherein A is the group CH (OH) - (v) from the formed compound of formula VII, the protection is removed by a cation with a strong acid to obtain an addition salt of a compound of formula I wherein A is a simple bond, CH 2, CHF, CH (OCH 3) or CH (OH), and (vi) said compound of formula I, if necessary, is obtained in the form of a free base by a reaction with a strong base and then other addition salts are obtained from said free base, or in step i) of process alternative C the NH 2 terminal end of a base of formula X is reacted R 1 is a protecting group, as stated above, and n is 6 · 25 or 8, with alkyl chloroformate or a symmetrical carbonate in an inert solvent at room temperature (15-25 ° C), (1) · (ii) the compound formed is aminolysed with an amine of formula VI (CH2) 4. R 2 represents the same as above, to obtain a compound of formula VII (NH 2) R 2 (CH 2) ·. JL. J1 (CH2) 6. R 2 and VII are the same as above and A is a CH 2 NH group, (iii) from the compound formed with the formula VII is removed by a reaction with a strong acid to obtain an addition salt of a compound of formula I wherein A is CH 2 NH, and (iv) said compound of formula I is obtained as needed in the form of a free base through a reaction with a strong base and then other addition salts are obtained from said free base, or in step (i) of the process alternative D, a compound of formula VI is (ch 2), Wherein R 3 is a protecting group as set forth above, with a carbonate of formula III1: C6H5 ° OCH2 ·. Wherein R2 is a linear or branched C1-C3 alkyl group or a phenylmethyl group, in an inert organic solvent at the refluxing temperature of the reaction mixture, when the molar ratio of the compounds is 1 mole of a compound of formula VI of a compound of formula III1, to obtain a compound of formula VIII, ____ (^ - ^ 2) t, v. 0. (NH) R 1 and R 2 are the same as above and A is a CH 2 group, (ii) from the formed compound of formula VIII, the protection is removed in a organic solvent in the presence of a strong base or by catalytic hydrogenation to obtain a compound of Formula IX 10. ou ^ (ch2) a ^ (CH2) 3 (IX) HO NH ^ NH * 1 where Rx denotes the same as above and A is CH 2 O, (iii) the formed compound of formula IX is condensed with an amine of formula II R 1 NH 20, 11, where R 1 is the same as above and n is 6 or 8, in an organic solvent in the presence of the carboxy group An activator and a nucleophilic agent at a temperature of 0 ° C - 40 ° C when the mole ratio of the compounds is 1; mole of a compound of formula IX / 1 mole of a compound: V; of formula II, to obtain a compound of formula VII: R 1 '(NH 2 O') R 1 (ch 2) j II II - (CH 2). (-R, r, (at 35 where Rx and n are the same as above and A is CH 2 O) (iv) from the compound of formula VII, the protection is removed by a reaction with a strong acid to obtain a Addition salt of a compound of formula I wherein A is CH 2 O and (v) said compound of formula I is obtained as needed in the form of a free base by a reaction with a strong base and then other addition salts are obtained from said free base, or in process option E, protection is removed from a compound of formula VII 10 R, 1 'NH oo R 1, (ch 2). il il ^ <ch2) 4 ^ (ch2) 3. In (vh) where n and A are the same as above and Rx is an alkoxycarbonyl-type protecting group by removing the protecting group by a reaction with a strong acid to substitute the residue Rx with H. 2. A process according to claim 1, characterized in that [2 - [[[4 - [[3- (amino) propyl] amino] butyl] amino] carbonyloxy] - is prepared. N- [6 - [(aminoiminomethyl) amino] hexyl] acetamide or its addition salt with a mineral acid or an organic acid. 25 Process according to claim 1, characterized in that N- [4 - [[3- (amino) propyl] amino] butyl] -N '- [6 - [(amino-iminomethyl) amino] hexyl] propanediamide is prepared. or its addition salt with a mineral acid or an organic acid. 30 4. An intermediate belonging to the synthesis of compounds of formula I, prepared by the process according to claim 1, characterized in that it is a compound of formula VII R oo N NH 2 n 2 -NH 4 NH c 2 NH R, (VII) 59 where Rx is an amino protecting group, n is 6 or 8 and A is a simple bond, CHα, CH (OH) -, CHF- , CH (OCH3 -), CH2NH, CH2O or CH (OCH2C6H5) group. »» I *